Journal article
ATP and AMP Mutually Influence Their Interaction with the ATP-binding Cassette (ABC) Adenylate Kinase Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) at Separate Binding Sites
The Journal of biological chemistry, Vol.288(38), pp.27692-27701
09/20/2013
DOI: 10.1074/jbc.M113.479675
PMCID: PMC3779764
PMID: 23921386
Abstract
Background:
Cystic fibrosis transmembrane conductance regulator (CFTR) has adenylate kinase activity (ATP + AMP ⇆ 2 ADP).
Results:
ATP enables CFTR photolabeling by 8-N
3
-AMP, and AMP increases 8-N
3
-ATP photolabeling at ATP-binding site 2.
Conclusion:
AMP interacts with CFTR in an ATP-dependent manner and alters ATP interaction with the adenylate kinase active center ATP-binding site.
Significance:
These findings exemplify nucleotide interactions with an ABC adenylate kinase.
Cystic fibrosis transmembrane conductance regulator (CFTR) is an anion channel in the ATP-binding cassette (ABC) transporter protein family. In the presence of ATP and physiologically relevant concentrations of AMP, CFTR exhibits adenylate kinase activity (ATP + AMP ⇆ 2 ADP). Previous studies suggested that the interaction of nucleotide triphosphate with CFTR at ATP-binding site 2 is required for this activity. Two other ABC proteins, Rad50 and a structural maintenance of chromosome protein, also have adenylate kinase activity. All three ABC adenylate kinases bind and hydrolyze ATP in the absence of other nucleotides. However, little is known about how an ABC adenylate kinase interacts with ATP and AMP when both are present. Based on data from non-ABC adenylate kinases, we hypothesized that ATP and AMP mutually influence their interaction with CFTR at separate binding sites. We further hypothesized that only one of the two CFTR ATP-binding sites is involved in the adenylate kinase reaction. We found that 8-azidoadenosine 5′-triphosphate (8-N
3
-ATP) and 8-azidoadenosine 5′-monophosphate (8-N
3
-AMP) photolabeled separate sites in CFTR. Labeling of the AMP-binding site with 8-N
3
-AMP required the presence of ATP. Conversely, AMP enhanced photolabeling with 8-N
3
-ATP at ATP-binding site 2. The adenylate kinase active center probe P
1
,P
5
-di(adenosine-5′) pentaphosphate interacted simultaneously with an AMP-binding site and ATP-binding site 2. These results show that ATP and AMP interact with separate binding sites but mutually influence their interaction with the ABC adenylate kinase CFTR. They further indicate that the active center of the adenylate kinase comprises ATP-binding site 2.
Details
- Title: Subtitle
- ATP and AMP Mutually Influence Their Interaction with the ATP-binding Cassette (ABC) Adenylate Kinase Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) at Separate Binding Sites
- Creators
- Christoph O Randak - From the Departments ofQian Dong - From the Departments ofAmanda R Ver Heul - From the Departments ofAdrian H Elcock - BiochemistryMichael J Welsh - Internal Medicine, and
- Resource Type
- Journal article
- Publication Details
- The Journal of biological chemistry, Vol.288(38), pp.27692-27701
- DOI
- 10.1074/jbc.M113.479675
- PMID
- 23921386
- PMCID
- PMC3779764
- NLM abbreviation
- J Biol Chem
- ISSN
- 0021-9258
- eISSN
- 1083-351X
- Publisher
- American Society for Biochemistry and Molecular Biology; 9650 Rockville Pike, Bethesda, MD 20814, U.S.A
- Grant note
- K08HL097071; HL51670; HL091842 / National Institutes of Health
- Alternative title
- Nucleotide Interactions with the ABC Adenylate Kinase CFTR
- Language
- English
- Date published
- 09/20/2013
- Academic Unit
- Neurology; Molecular Physiology and Biophysics; Pulmonary, Critical Care, and Occupational Medicine; Pulmonary Medicine; Stead Family Department of Pediatrics; Physics and Astronomy; Biochemistry and Molecular Biology; Neurosurgery; Internal Medicine
- Record Identifier
- 9984020192702771
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